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Gene transfer systems for phytopathogenic corynebacteria have not been reported previously. In this paper we describe a conjugative 46-megadalton plasmid (pDG101) found in Corynebacterium flaccumfaciens subsp. oortii COIOI that mediates resistance to arsenite, arsenate, and antimony(III). Transfer of the plasmid from CO101 to four other strains from the C. flaccumfaciens group occurred between cells immobilized on nitrocellulose filters or on agar surfaces. Transconjugant strains expressed the same levels of metal resistance as the donor strain and were able to act as donor strains in subsequent matings. The physical presence of the plasmid was detected by agarose gel electrophoresis. Arsenite-sensitive derivatives of the donor and transconjugant strains were obtained after heat treatment; these were cured of pDG101.